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J. Biol. Chem., Vol. 266, Issue 36, 24276-24286, 12, 1991
BL Bassler, C Yu, YC Lee and S Roseman
Chemotaxis of the marine bacterium Vibrio furnissii to chitin
oligosaccharides has been described (Bassler, B. L., Gibbons, P. J., Yu,
C., and Roseman, S. (1991) J. Biol. Chem. 266, 24268-24275). Some steps in
catabolism of the oligosaccharides are reported here. GlcNAc, (GlcNAc)2,
and (GlcNAc)3 are very rapidly consumed by intact cells, about 320 nmol of
GlcNAc equivalents/min/mg of protein. (GlcNAc)4 is utilized somewhat more
slowly. During these processes, there is virtually no release of hydrolysis
products by the cells. The oligosaccharides enter the periplasmic space
(via specific porins?) and are hydrolyzed by a unique membrane-bound
endoenzyme (chitodextrinase) and an exoenzyme
(N-acetyl-beta-glucosaminidase; beta-Glc-NAcidase). The genes encoding
these enzymes have been cloned and expressed in Escherichia coli. The
chitodextrinase cleaves soluble oligomers, but not chitin, to the di- and
trisaccharides, while the periplasmic beta- GlcNAcidase hydrolyzes the
GlcNAc termini from the oligomers. The end products in the periplasm,
GlcNAc and (GlcNAc)2 (possibly (GlcNAc)3) are catabolized as follows. (a)
Disaccharide pathway, A (GlcNAc)2 permease is apparently expressed by
Vibrio furnissii. Translocated (GlcNAc)2 is rapidly hydrolyzed by a
soluble, cytosolic beta- GlcNAcidase, and the GlcNAc is phosphorylated by
an ATP-dependent, constitutive kinase to GlcNAc-6-P. (b) Monosaccharide
pathway, Periplasmic GlcNAc is taken up by Enzyme IINag of the
phosphoenolpyruvate:glycose phosphotransferase system, yielding GlcNAc-
6-P, the common intermediate for both pathways. Finally, GlcNAc-6-P---- Ac-
+ GlcNH2-6-P----Fru-6-P + NH3. (GlcNAc)2 is probably the "true" inducer of
the chitin degradative enzymes described in this report and, depending on
its concentration in the growth medium, differentially induces the
periplasmic and cytosolic beta-GlcNAcidases. The disaccharide pathway
appears to be the most important when the cells are confronted with low
concentrations of the oligomers (e.g. in chemotaxis swarm plates). The
relative activities of the induced enzymes suggest that the rate-limiting
steps in oligosaccharide catabolism are the glycosidase activities in the
periplasm.
Chitin utilization by marine bacteria. Degradation and catabolism of chitin oligosaccharides by Vibrio furnissii
McCollum-Pratt Institute, Johns Hopkins University, Baltimore, Maryland 21218.
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